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Please use this identifier to cite or link to this item: http://hdl.handle.net/123456789/17071

Title: New highly sensitive enzyme immunoassay for the determination of pravastatin in human plasma.
Authors: Ibrahim A Darwish
Keywords: Antibodies, Anticholesteremic Agents, Binding, Competitive, Horseradish Peroxidase, Humans, Immunoenzyme Techniques, Pravastatin, Rabbits, blood, methods
Issue Date: 2009
Publisher: Talanta
Abstract: New highly sensitive enzyme immunoassay (EIA) has been developed and validated for the determination of pravastatin (PRV) in human plasma samples. PRV was coupled to keyhole limpt hemocyanin (KLH) and bovine serum albumin (BSA) via its terminal carboxylic acid group by carbodiimide reagent. PRV-KLH conjugate was used as an immunogen for raising anti-PRV polyclonal antibody in rabbits. The generated anti-PRV antibody recognized PRV with high affinity and selectivity. PRV-BSA conjugate was immobilized onto microwell plates and used as a solid phase. The assay involved a competitive binding reaction between PRV, in plasma sample, and the immobilized PRV-BSA for the binding sites on a limited amount of the anti-PRV antibody. The anti-PRV antibody bound to the plate wells was quantified with horseradish peroxidase-labeled anti-immunoglobulin second anti-rabbit IgG antibody and 3,3',5,5'-tetramethylbenzidine as a substrate for the peroxidase enzyme. The concentration of PRV in the sample was quantified by its ability to inhibit the binding of the anti-PRV antibody to the immobilized PRV-BSA and subsequently the color development in the assay wells. The conditions of the proposed EIA were investigated and the optimum conditions were employed in the determination of PRV in plasma samples. The assay limit of detection was 0.2 ng mL(-1) and the effective working range at relative standard deviation (RSD)
URI: http://hdl.handle.net/123456789/17071
Appears in Collections:College of Pharmacy

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